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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is breast carcinoma.
Cyclins bind to and regulate the activity of the Cyclin dependent protein kinases (CDKs). Cyclin B1 or CCNB1 is a regulatory protein involved in mitosis. It is essential for the control of the cell cycle at the G2/M transition. Cyclin B1 complexes with p34 (cdc2) to form the maturation-promoting factor (MPF). Two alternative transcripts have been found, a constitutively expressed transcript and a cell cycle-regulated transcript, that is expressed predominantly during G2/M phase. Cyclin B1 is not ubiquitinated during G2/M phase, resulting in its steady accumulation during G2 phase, followed by abrupt APC dependent destruction at the end of mitosis. Destruction of Cyclin B1 is required for cell cycle progression. Cyclin B1 is overexpressed in various cancers, including breast, prostate, and non-small cell lung cancer.
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